Revised Project Summary/Abstract The overall goal of the proposed research is to identify epigenetic changes in biological pathways associated with protection from Type 2 Diabetes (T2D) in Yup?ik Alaska Native people consuming a traditional diet. The Yup?ik traditional dietary pattern is characterized by high protein, high fat, and low carbohydrate intakes, as well as n-3 polyunsaturated fatty acid levels that are up to 20 times higher than in the general U.S. population. High levels of traditional food intake in Yup?ik people are positively associated with adiponectin and inversely associated with c-reactive protein, triglycerides, and the leptin:adiponectin ratio, suggestive of increased insulin sensitivity. However, the diet of Yup?ik people is transitioning to increased consumption of highly processed market foods, and the prevalence of overweight and obesity is beginning to mirror that of the general U.S. population. Even so, the prevalence of metabolic syndrome (15%) and T2D (3%) among Yup?ik people remains lower than in the general U.S. population. Adherence to a Yup?ik traditional dietary pattern is likely to be protective in Yup?ik people, although the mechanisms of protection are not well understood. We will use the nitrogen stable isotope ratio (delta[15]N) as a validated, objective measure of the traditional dietary pattern to explore these mechanisms. Although considerable advances have been made in the identification of genetic factors contributing to metabolic diseases, less than 10% of the heritability of T2D phenotypes is explained by known genetic variation. Epigenetic mechanisms have recently been proposed as a link between genetic risk and environmental exposures such as diet, thereby providing a mechanism for the modification of genetic predisposition and insight into 90% of the heritability that is not explained. The originally proposed research had 4 specific aims: 1) use Illumina MethylationEPIC microarrays to comprehensively screen >850,000 methylation sites on stored peripheral blood DNA samples from 569 Yup?ik people at the highest and lowest deciles of traditional dietary intake; 2) fine map methylation sites by bisulfite sequencing the 20 most promising candidate gene regions in 192 stored Yup?ik DNA samples and determine the generalizability of the findings in 192 stored Greenlandic Inuit DNA samples; 3) identify dynamic, metabolically-related epigenetic variation within individuals by performing longitudinal epigenetic analyses on 150 new peripheral blood DNA samples derived from a subset of the 576 Yup?ik participants in Aim 1; and 4) perform functional analyses in newly collected RNA samples from 192 past participants at the extremes of traditional food intake to determine the downstream changes in gene expression associated with dietary pattern and altered genomic methylation. Aims 2 and 4 (listed above) were dropped from this grant because we received a reduced budget from NIDDK.